应用荧光定量RT-PCR方法检测PRRSV变异株在仔猪体内动态分布  

The dynamic distribution of PRRSV mutant strain in infected piglets by the fluorescent quantitative RT-PCR

作　　者：荣福龙[1,2] 刘永刚[1] 孙广力[1,2] 王淑杰[1] 王洪峰[1] 石文达[1] 李丽琴[1] 徐明明[1,2] 孙刚 田志军[1] 蔡雪辉[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/猪传染病研究室,黑龙江哈尔滨150001 [2]东北农业大学,黑龙江哈尔滨150001 [3]黑龙江省动物卫生监督所,黑龙江哈尔滨150090

出　　处：《中国预防兽医学报》2010年第9期681-686,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：中央基本科研业务费(2008-01)

摘　　要：为进一步了解高致病性猪繁殖与呼吸综合征病毒(PRRSV)变异株(HuN4株)在体内的动态分布规律和特点,探讨病毒对组织器官的嗜性,本研究将HuN4株和细胞传代致弱毒株HuN4-F65人工感染40日龄健康仔猪,分别于感染后1d、3d、5d、7d、10d、14d、21d和28d各迫杀2头,并应用建立的荧光定量RT-PCR方法对脑、颌下淋巴结、扁桃体、肺脏、肝脏和十二指肠等组织及血清中的PRRSV进行定量检测。结果显示:感染后HuN4组各器官病毒载量比HuN4-F65组高100倍,病毒载量峰值期出现在感染后7d,随后呈下降趋势。HuN4株在仔猪体内分布广泛,在21d的试验期内持续存在,并且各器官的病毒载量呈正态分布规律;而HuN4-F65株血清病毒载量较低,并且集中在单核巨噬细胞相对密集的扁桃体和颌下淋巴结,与HuN4株主要嗜性器官肺脏相比发生了改变。To understand the kinetics and distribution of highly pathogenic porcine reproductive and respiratory syndrome virus （PRRSV） mutant strain （HuN4 strain） in piglets,forty-day-old healthy piglets were inoculated with the virulent HuN4 strain or the attenuated strain HuN4-F65,and slaughtered at 3,5,7,10,14,21 and 28 days post-inoculation （dpi）,respectively. The brain,submandibular lymph node,tonsil,lung,liver,duodenum and serum samples were collected and detected by real-time RT-PCR. The results showed that the viral load in HuN4 infected group was significantly higher than that in HuN4-F65 group,which peaked on 7 dpi,and decreased at the later stage. The HuN4 strain was widely distributed during entire 21 dpi in various organs,especially in the lung. In contrast,the serum viral load in HuN4-F65 group appeared lower,and mainly in the tonsil and submandibular lymph nodes where monocyte-macrophage cells are relatively dense. These results indicated that the HuN4 viral tropism to the lung and other organs had sustained significant changes after attenuation.

关 键 词：PRRSV变异株 荧光定量RT-PCR 抗原动态分布 

分 类 号：S852.65[农业科学—基础兽医学]